In this post we intend to give an example of protocol to carry out experiments of toxicity in C.elegans.
Steps to collect worms:
1- Wash agar plate synchronized in L4 or adults with buffer or water.
2- Generate a suspension of 50 worms / 100 ul of medium. The total volume to be prepared will depend on the assay design (number of wells to be used).
3- Place in 96-well plate, 100 ul of the worm suspension per well (50 worms per well).
4- Seal the plate with parafilm and let stand for 45 min.
5- Configure the experimental groups (one per toxic concentration) in the WMicrotracker software.
6- Place the plate in the WMicrotracker and record zero time activity (basal activity) for 30 minutes.
7- Place 10 ul of toxic solution (minimum 4 different concentrations) and re-seal the plate. Use at least 4 wells per concentration (technical repetitions).
8- Place the plate in the WMicrotracker and record the activity for 120 minutes.
9- Graph the activity in blocks of 30 minutes, basal activity vs. toxic concentration.